Slack(KCNT1)通道癫痫相关突变体在额叶癫痫发作中作用机制的研究

项目名称： Slack(KCNT1)通道癫痫相关突变体在额叶癫痫发作中作用机制的研究

项目编号： No.81471314

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 张赭

作者单位： 徐州医科大学

项目金额： 70万元

中文摘要： 近来,遗传学的研究发现几种家族性遗传癫痫患者携带Slack钾离子通道基因的突变。然而这些突变如何改变Slack通道的功能和在癫痫发作中的作用都未得到阐明。本课题以此为切入点，运用爪蟾卵母细胞表达系统，采用内面向外的膜片钳和双电极记录的电生理记录方法研究揭示这些突变体通道生物物理特性的改变；并结合结晶学、分子模拟及其它生化学方法阐明这些突变体生物物理特性改变的构象基础和机制。同时，本研究也将采用在体电生理技术、免疫双标等方法以及基因工程小鼠的动物模型，包括Slack通道基因敲除的小鼠和Gain-of-function的Slack通道突变体的小鼠， 运用海人藻酸诱导的癫痫模型，进一步验证和阐释Slack通道突变体在癫痫发作中的作用和其机制。本课题将为深入理解Slack通道激活机制提供理论基础，也为临床医生判断这些突变在癫痫发病中的作用以及寻求相应的治疗方法和药物提供依据。

中文关键词： 离子通道；癫痫发作；钾通道；神经电生理

英文摘要： Recently， genetic research revealed that several kinds of genetical family epilepsy patients carreid Slack channel mutations.However, how the mutants changed Slack channel function and their role in focal seizures remains elusive.This project will take advantage of Xneopus expression system and use inside-out patch and two-electrode voltage clamp method to reveal how the mutants change the biophysical property of Slack channels. In addition, we are going to combine crystalization, molecular simulation and other biochemical methods to address how the Slack mutant channel conformational change and the underlying mechanism of biophysical property change.In the meantime, our project will also set up kanatic acid induced epilepsy model on both Slack gene knockout and Gain-of-function mice model to test the role of Slack channel mutants in focal seizures by using brain-slice recording , immunostaining and whole cell recording technique.This project is going to provide a solid base for further understanding the Slack channel activation mechanism. Also, the results of this project can help clininal doctor judge the role of these mutants on epilepsy patients who carried these mutants and search for effective treatment method and drugs.

英文关键词： ion channels;epilesy seizure;potassium channel;neuroelectrical physiolog